Tolerability and safety of the intake of bovine milk oligosaccharides extracted from cheese whey in healthy human adults

Jennifer T Smilowitz, Danielle G Lemay, Karen M Kalanetra, Elizabeth L Chin, Angela M Zivkovic, Melissa A Breck, J Bruce German, David A Mills, Carolyn Slupsky, Daniela Barile, Jennifer T Smilowitz, Danielle G Lemay, Karen M Kalanetra, Elizabeth L Chin, Angela M Zivkovic, Melissa A Breck, J Bruce German, David A Mills, Carolyn Slupsky, Daniela Barile

Abstract

Mechanistic research suggests a unique evolutionary relationship between complex milk oligosaccharides and cognate bifidobacteria enriched in breast-fed infants. Bovine milk oligosaccharides (BMO) were recently identified as structurally and functionally similar to human milk oligosaccharides. The present single-blind three-way crossover study is the first to determine the safety and tolerability of BMO consumption by healthy human participants (n 12) and its effects on faecal microbiota and microbial metabolism. Participants consumed each supplement (placebo-control; low- and high-BMO doses) for eleven consecutive days, followed by a 2-week washout period prior to initiating the next supplement arm. Low and high BMO doses were consumed as 25 and 35 % of each individual's daily fibre intake, respectively. Safety and tolerability were measured using standardised questionnaires on gut and stomach discomfort and stool consistency. Faecal extracts were profiled for bacterial populations by next-generation sequencing (NGS) and bifidobacteria presence was confirmed using quantitative PCR. Urine was analysed for changes in microbial metabolism using nuclear magnetic resonance spectroscopy (1H-NMR). Consumption of both the low and high BMO doses was well tolerated and did not change stool consistency from baseline. Multivariate analysis of the NGS results demonstrated no change in faecal microbiota phyla among the placebo-control and BMO supplement groups. In conclusion, BMO supplementation was well tolerated in healthy adults and has the potential to shift faecal microbiota toward beneficial strains as part of a synbiotic treatment with probiotic cultures that selectively metabolise oligosaccharides.

Keywords: BMO, bovine milk oligosaccharides; Bovine milk oligosaccharides; GI, gastrointestinal; Gastrointestinal tolerability; HMO, human milk oligosaccharides; HNC, Human Nutrition Center; Mass spectrometry; Microbiota; Next-generation sequencing; OTU, operational taxonomic unit.

Figures

Fig. 1.
Fig. 1.
Self-report stool consistency levels by participants during the run-in (□) and intervention (░) period by each study participant. Values are means (n 9 per supplemental arm), with standard deviations represented by vertical bars. BMO, bovine milk oligosaccharides.
Fig. 2.
Fig. 2.
Faecal relative abundance of the family Peptostreptococcaceae in each participant in response to the placebo-control and high-bovine milk oligosaccharide (BMO) supplement arms on days 0 (), 4 () and 11 (■) (n 9 per supplemental arm). The linear discriminant analysis effect size between the high-BMO and placebo-control groups for the faecal relative abundance of the family Peptostreptococcaceae was 3·4 (P < 0·05).
Fig. 3.
Fig. 3.
Faecal bifidobacteria levels at day 0 (□), day 4 () and day 11 (░) in response to the placebo-control (PC), low-bovine milk oligosaccharide (LB) and high-bovine milk oligosaccharide (HB) supplement arms. Values are means (n 9 per supplemental arm), with standard deviations represented by vertical bars.
Fig. 4.
Fig. 4.
Principal components (PC) analysis of urine metabolite concentrations in different treatment arms. (A) Comparison of days 4 and 11 in the placebo-control (○), low-dose (), and high-dose (●) bovine milk oligosaccharide (BMO) groups. The variances along PC1 and PC2 were 42 and 5 %, respectively. (B) Mean PC2 value for each of seven subjects who provided a urine sample at all time points during all three treatment arms in the study, with standard errors for days 4 and 11.
Fig. 5.
Fig. 5.
Percentage change between day 11 and day 0 for urinary cis-aconitate, myo-inositol, 4-hydroxyphenylacetate, 2-oxoglutarate and mannitol across all three supplement arms: placebo-control (□), low-bovine milk oligosaccharide (BMO) () and high-BMO (■). Values are means (n 7 per supplemental arm), with standard deviations represented by vertical bars. The mean percentage change for urinary cis-aconitate was significantly different in response to the high-BMO compared with the placebo-control (P < 0·05).

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